Subject Number : S-20-KU-0003
Support Type : Common use (including technical support necessary for the training)
Proposal Title (English) : Determination of specific lipid to membrane protein using self-assembled monolayer gold nanoparticle
Username (English) : Supakorn Wangamnuayporn, Masanao Kinoshita,Takayuki Kawai ,Nobuaki Matsumori
Affiliation (English) : Graduate School of Science, Kyushu University

1. Summary
In this study, self-assembled monolayer (SAM)-modified gold nanoparticle (AuNP) is developed for efficient determination of membrane protein (MP)-specific lipids on the lipid mixture, based on the SAM-coated gold sensor chip idea. 1 Bacteriorhodopsin (bR), a seven-transmembrane MP within purple membrane, was immobilized on the functionalized AuNP to investigate lipid-protein interaction. The result showed that S-TGA-1 which was reported to have strong affinity to bR are successfully detected after screening lipid sample with bR-immobilized AuNPs.

2. Experimental
In this study, AuNPs was coated with the C6-SAM, following by bR immobilization. The MP-bound lipid species was quantified by MALDI-TOF/MS after introducing lipid sample from the purple membrane.

3. Results and Discussion
After introducing lipid mixture from purple membrane, bR-immobilized AuNPs successfully detected S-TGA-1 as bR-specific lipid, consistent with the result from the SPR-based method. 2 MALDI-TOF/MS spectra showed higher peak ratio of S-TGA-1, comparing with other lipid species in the lipid sample, after lipid screening by bR-immobilized AuNPs.

4. Others
Reference:
(1) M. Inada, M. Kinoshita, A. Sumino, S. Oiki, N. Matsumori, Anal. Chim. Acta 2019, 1059, 103-112.
(2) M. Inada, M. Kinoshita, N. Matsumori, ACS Chem Biol. 2020, 15(1), 197-204.

5. Publication/Presentation
(1) Supakorn Wangamnuayporn, Masanao Kinoshita, Nobuaki Matsumori， 日本化学会 第101春季年会，2020/03/20
(2) Supakorn Wangamnuayporn, Masanao Kinoshita, Nobuaki Matsumori， 第14回バイオ関連化学シンポジウム，2020/09/08

6. Patent
N/A